RNA sequencing (RNA-seq) is a widely used method employing next generation sequencing (NGS) to discover and quantify the abundance of RNA molecules in biological samples.Why do we do RNA-seq? RNA-seq has revolutionized approach to transcriptomics, it allows high-throughput being precise and cos...
To this end, workflow standardization from sample collection and storage through nucleic acid purification and next-generation sequencing (NGS) library preparation is critical in order to compare and combine separate studies in consortium-based approaches at a global scale. Eppendorf, a leading life sci...
although alignment-independent methods can also be used. Raw read data should be quality-controlled with tools, such as FASTX-toolkit [56], FASTQC [57] or NGS QC [
Standard pipelines for NGS analysis.aAlignment and pre-processing of NGS data for an individual sample. Raw sequence data in FASTQ format are aligned to the reference sequence, with the resulting alignments typically stored in binary alignment/map (BAM) file format. Marking of duplicates in the B...
While years ago the costs associ- ated with this methodology were high, the drop in prices for sequencing has opened the door for the application of NGS to biomarker discovery and as a screening tool. To adapt to this new field, multiple protocols for library preparation have been developed ...